Genetic Analysis of the ts-Lethal Mutant / Reveals Its Role in Cell Morphology and Oxidative Phosphorylation in .

Pa0665 in shares homologous sequences with that of the essential A-type iron-sulfur (Fe-S) cluster insertion protein ErpA in . However, its essentiality in and its complementation with has not been experimentally examined. To fulfill this task, we constructed plasmid-based -mutant / using a three-step protocol. The mutant displayed growth defects at 42 °C, which were complemented by expressing . Microscopic observations indicated a petite cell phenotype for / at 42 °C, correlated with the downregulation of the gene. RNA sequencing revealed significant transcriptional changes in genes associated with the oxidative phosphorylation (OXPHOS) system, aligning with reduced ATP levels in / under 42 °C. Additionally, the -mutant showed heightened sensitivity to HO at 42 °C. Overall, our study demonstrates the essential role of for OXPHOS function and is complemented by . We propose that the plasmid-based -allele is useful for genetic analysis of essential genes of interest in .